WO2014076342A1 - Méthode d'obtention de données utiles pour le diagnostic, le pronostic et la prédiction d'une réponse au traitement de l'adénocarcinome du pancréas - Google Patents

Méthode d'obtention de données utiles pour le diagnostic, le pronostic et la prédiction d'une réponse au traitement de l'adénocarcinome du pancréas Download PDF

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WO2014076342A1
WO2014076342A1 PCT/ES2013/070786 ES2013070786W WO2014076342A1 WO 2014076342 A1 WO2014076342 A1 WO 2014076342A1 ES 2013070786 W ES2013070786 W ES 2013070786W WO 2014076342 A1 WO2014076342 A1 WO 2014076342A1
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seq
genes
vnn1
ankrd22
clec4d
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PCT/ES2013/070786
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Spanish (es)
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Juan Ramón DELGADO PÉREZ
Antonia ARÁNEGA JIMÉNEZ
José Carlos PRADOS SALAZAR
Octavio CABA PÉREZ
Consolación MELGUIZO ALONSO
Fernando RODRÍGUEZ SERRANO
Raul Ortiz Quesada
María Celia VÉLEZ FERNÁNDEZ
Ignacio ROJAS RUIZ
Alberto Prieto Espinosa
Javier PÉREZ FLORIDO
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Universidad De Granada
Servicio Andaluz De Salud
Universidad De Jaén
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Publication of WO2014076342A1 publication Critical patent/WO2014076342A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/16—Primer sets for multiplex assays
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis

Definitions

  • the present invention is within medicine and molecular biology, and refers to a method of obtaining useful data for the early diagnosis of pancreatic adenocarcinoma, as well as to evaluate the response to the treatment of said disease, which allows the establishment of an individual pattern of recognition (quantitative) specific, which is modified post-treatment, allowing the establishment of groups of patients.
  • Pancreatic adenocarcinoma ⁇ Pancreatic Ductal Adenocarcinoma PDAC is an epithelial neoplasm of pancreatic ducts (Kloppel et al., 1996) and the fourth leading cause of cancer death in the world (Sharma et al., 201 1, JOP 12 (4): 343-6).
  • PDAC has the worst prognosis of all gastrointestinal tumors, with a mortality rate very close to the incidence (Karanjawala et al., 2008, Am J Surg Pathol; 32: 188-96).
  • the current survival rate of patients affected by PDAC is less than 5% (Sharma et al., 201 1, JOP 12 (4): 343-6; Huang et al., 2010, Am J Gastroenterol 105 (7): 1661-9).
  • pancreatic tumor cannot be resected when there are distant metastases, (eg liver, extra-abdominal, peritoneum, lymph nodes) (Li et al., 2004) or there is a locally advanced tumor.
  • distant metastases eg liver, extra-abdominal, peritoneum, lymph nodes
  • a locally advanced tumor e.g., a tumor of the superior mesenteric artery, the inferior vena cava, the aorta, the celiac trunk or the superior mesenteric vein and the portal venous system.
  • CA19-9 or Lewis sialylated antigen (Buxbaum et al., 2010; JOP 1 1 (6): 536-44.), With an average sensitivity of 80% and specificity of 90% (Steinberg et al. ., 1990; Am J Gastroenterol; 85: 350-5).
  • CA19-9 may be useful for monitoring treatment and determining whether the disease may be recurrent (Locker et al., 2006, J. Clin. Oncol ;.
  • CA19-9 levels are not sufficient to identify primary stages of PDACs, and only 65% of patients with resectable pancreatic cancer have elevated levels of this antigen (Goggins et al., 2005; J Clin Oncol; 23: 4524-31).
  • microarrays The global gene expression profile of microarrays provides important information about the molecular characteristics of cancer, and is widely used for the development of biomarkers.
  • PDAC the first tissue RNA gene expression profile that was based on microarrays was published in 2002 (Lacobuzio-Donahue et al., 2002; Am J Pathol 160 (4): 1239-49). Since then, many types of genes have been identified whose expression is deregulated in primary pancreatic cancer compared to normal pancreatic tissue, which can be very useful for the detection of pancreatic cancer, from its stages, or to predict its prognosis (Karanjawala et al., 2008, J Surg Pathol 32: 188-96).
  • Peripheral blood is an accessible source of cells with which to investigate important issues.
  • circulating cells can be viewed as scouts, continuously maintaining a thorough surveillance of signs of infection or other threats in the body (Whitney et ai, 2003. Proc Nati Acad Sci U S A 100 (4): 1896-901).
  • genetic expression in PBMCs ⁇ Peripheral Blood Mononuclear Cells is altered as if it were a disease-associated signature (Whitney et al., 2003; Proc. Nati. Acad. Sci.
  • peripheral blood circulating mononuclear cells can serve as a controller of the physiological state of the organism.
  • these cells pass through several tissues, their reaction with the medium is captured in a complex transcriptional response that is measured through a profile.
  • blood cells communicate with extracellular cells and matrices in almost all body tissues and organs (Liotta et ai., 2003; Nature; 425 (6961): 905).
  • the differential genetic expression profile of PBMCs is potentially a very useful tool for early diagnosis. This is especially important considering that two of the most likely mechanisms on which this differential expression is based would be the recognition and at the same time the evasion of cancer by the immune system. While other biomarkers such as CA19-9 are released from cancer cells and increase with the increase in tumor burden, differential expression in PBMCs could begin, at least partially, as soon as immune evasion or cancer's immunogenicity is established. It has been shown that evasion of the immune system begins before the disease is considered malignant in PDAC. Therefore, the analysis of differential genetic expression In immune cells it could offer the ability to detect neoplastic lesions even before it becomes invasive (Baine et al., 201 1. PLoS One. 201 1; 6 (2): e17014).
  • pancreatic cancer protein microarrays have been performed to detect serum markers (Orchekowski et al., 2005; Cancer f ⁇ es, 65: 1 1 193-202), as well as cDNA microarrays, to detect overexpressed genes in the neoplastic epithelium in 10 pancreatic cancers compared with 10 samples of non-tumor pancreas (Logsdon et al., 2003. Physe cancer 63: 2649-57).
  • pancreatic cancer Likewise, comparative studies of the microarray analyzes used in pancreatic cancer have been carried out in order to find genes and biomarkers that could be used in new diagnostic and therapeutic strategies (Brandt et al., 2004. Pancreatology A: 587-597).
  • the authors of the present invention have selected a group of genes whose expression correlates with early diagnosis, prognosis and response to the treatment of pancreatic adenocarcinoma.
  • the present invention provides a method of obtaining useful data for the early diagnosis of adenocarcinoma of the pancreas, as well as for evaluating the response to the treatment of said disease, allowing the establishment of groups of patients.
  • a first aspect of the invention relates to the use of any of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3, FAIM3 or any combination thereof, for diagnosis, for prognosis, or for the prediction of the response to the chemotherapy treatment of adenocarcinoma of the pancreas.
  • Another aspect of the invention relates to the simultaneous use of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3, FAIM3 genes for obtaining useful data in the diagnosis, prognosis or prediction of the response to chemotherapy treatment of pancreatic adenocarcinoma.
  • the independent use of any of them or any of their combinations could be sufficient for the diagnosis, prognosis or prediction of the response to the treatment of said disease.
  • Another aspect of the invention relates to a method of obtaining useful data for the diagnosis, prognosis or prediction of the response to chemotherapy treatment of adenocarcinoma of the pancreas, comprising:
  • step (c) compare the expression of the gene or the genes of step (b) with a reference amount.
  • Another aspect of the invention relates to a method of obtaining useful data for the diagnosis, prognosis or prediction of the response to chemotherapy treatment of adenocarcinoma of the pancreas, hereinafter the first method of the invention, comprising:
  • step (b) simultaneously detect the level of expression of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, FAIM3 genes in the isolated sample of (a), and (c) compare the expression of the genes of step (b) with a reference amount.
  • steps (b) and / or (c) of the methods described above can be totally or partially automated, for example, by means of a robotic sensor device for the detection of the quantity in step (b) or the computerized comparison in step (c).
  • Another aspect of the invention relates to a method for the diagnosis, prognosis or prediction of the response to the treatment of adenocarcinoma of the pancreas in an individual, hereafter referred to as the second method of the invention, comprising steps (a) - (c ) according to the first method of the invention, and further comprises:
  • step (d) diagnose the individual of step (a) as an individual with adenocarcinoma of the pancreas, when he has an increased expression of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, FAIM3 genes, in the sample obtained in (a), in relation to the amount of expression detected for said genes in a population of reference patients. More preferably, the expression of said genes is twice the basal expression in the reference population.
  • the isolated biological sample of an individual from step (a) is obtained from peripheral blood, and / or comprises peripheral blood cells ⁇ peripheral blood mononuclear cells PBMCs).
  • the studies of the authors of the present invention have allowed us to obtain information on gene expression patterns in patients with unresectable adenocarcinoma of the pancreas versus control subjects.
  • the study using computer techniques of the results obtained has allowed us to obtain a model of the modifications that are significant in this type of pathology.
  • pancreatic adenocarcinoma is unresectable.
  • the detection of gene expression can be performed by any means known in the state of the art.
  • the authors of the present invention have demonstrated that the detection of the expression in a quantitative way allows to differentiate between the patient with adenocarcinoma of the pancreas and the healthy individual.
  • the concentration measurement can be carried out directly or indirectly.
  • Direct measurement refers to the measure of gene expression, based on a signal that is obtained directly from the transcripts of said genes, or from the proteins to which they are translated, and that is directly correlated with the number of molecules of RNA or proteins produced by genes.
  • Said signal - which we can also refer to as an intensity signal - can be obtained, for example, by measuring an intensity value of a chemical or physical property of said products.
  • the indirect measurement includes the measurement obtained from a secondary component or a biological measurement system (for example the measurement of cellular responses, ligands, "tags" or enzymatic reaction products).
  • the term "comparison”, as used in the description, refers to, but is not limited to, the comparison of expression levels of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, FAIM3 genes in the biological sample to be analyzed, also called the biological problem sample, with the expression levels of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3, FAIM3 genes of one or more desirable reference samples described elsewhere of this description.
  • the reference sample can be analyzed, for example, simultaneously or consecutively, together with the problem biological sample.
  • the comparison described in section (c) of the method of the present invention can be performed manually or assisted by a computer.
  • Gene expression profile means the gene profile obtained after quantification of mRNA and / or protein produced by the genes of interest or biomarkers, that is, by the genes ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1 , F5, ⁇ RAK3 and FAIM3, in an isolated biological sample.
  • the gene expression profile is preferably performed by determining the level of mRNA derived from its transcription, after extracting the total RNA present in the isolated biological sample, which can be performed using protocols known in the state of the art.
  • the level of mRNA derived from the transcription of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3 and FAIM3 genes can be determined, for example, but not limited to, by amplification by polymerase chain reaction (PCR), retrotranscription in combination with the polymerase chain reaction (RT-PCR), quantitative RT-PCR, retrotranscription in combination with the ligase chain reaction (RT-LCR), or any other amplification method of nucleic acids; serial analysis of gene expression (SAGE, SuperSAGE); DNA chips made with oligonucleotides deposited by any mechanism; DNA microarrays made with oligonucleotides synthesized in situ by photolithography or by any other mechanism; in situ hybridization using specific probes labeled with
  • the gene expression profile could also be obtained by the detection and / or quantification of the proteins resulting from the translation of the mRNA derived from the transcription of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3 and FAIM3 genes. for example, but not limited to, western blot immunodetection.
  • Quantitative detection of the expression of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3 and FAIM3 genes can be more preferably performed by real-time PCR (RT-PCR or RTqPCR).
  • the real-time detection of the amplified products can be carried out by means of the use of fluorescent molecules that are intercalated in the double-stranded DNA or by hybridization with different types of probes.
  • the detection of the expression levels of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3, FAIM3 genes is performed by Q-RT-PCR.
  • Quantitative real-time PCR is a technique for quantifying sensitive and reproducible gene expression that can be used in particular for the expression of the gene profile in cells and tissues. Any procedure may be used for the evaluation of the results of the RT-PCR and the AACt procedure may be preferred.
  • the AACt procedure will involve a "control sample” and a "subject sample”.
  • the "subject sample” is a sample from the subject to be analyzed.
  • a target gene here: the gene of interest
  • an endogenous control gene as described below
  • the efficiency of PCR amplification can be defined as the percentage of amplification (from 0 to 1).
  • software normally measures the number of cycles of each sample in which the fluorescence crosses an arbitrary line (PCR amplification indicator), the threshold. This crossing point is the Ct value. More diluted samples will cross to subsequent Ct values.
  • the Ct of a nucleic acid from the gene of interest is divided by the Ct of the nucleic acid from the endogenous control in the same sample to normalize the variation in the quantity and quality of RNA between different samples and obtain the relative expression (with respect to the endogenous control) of each of the "sample of the subject" and of the "control sample”.
  • this is carried out in duplicate, triplicate, quadruplicate and similarly, respectively.
  • An ACt value of the control can be adequately obtained by calculating the average of the ACt values obtained from samples of a control group of several individuals with which the values of the "subject sample" are to be compared.
  • the control group (from which the average value is calculated) consists of individuals suitable for the respective purposes (for comparison).
  • the skilled person will learn from this disclosure that a suitable control group is for a specific purpose.
  • the present invention can be practiced by omitting the determination of the ACt value of the control group, that is, determining (only) the ACt value of the "subject sample” and then comparing this with the respective one. average ACt value of the control indicated in the examples.
  • the detection of the expression product of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, FAIM3 genes is performed by Northern Blot Transfer.
  • the detection of the gene expression product ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, FAIM3 is performed using microarrays.
  • the ANKRD22 or nkyr ⁇ n repeat domain-containing protein 22 gene is found on chromosome 10.
  • ANKRD22 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 1, and which would comprise various variants from:
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 1,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 1, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the ANKRD22 protein.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 2.
  • the ARG1 or arginase gene, liver, (arginase-1; liver-type arginase; type I arginase) is found on chromosome 6 (6q23).
  • ARG1 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 3, and which would comprise various variants from: a) acid molecules nucleic encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 3,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, a 98% or 99% with SEQ ID NO: 3, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the ARG1 protein.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 4.
  • the S100A8 or S100 calcium binding protein A8 gene (60B8AG, CAGA, CFAG, CGLA, CP-10, L1 Ag, MA387, MIF, MRP8, NIF, P8, RP-8; S100 calcium-binding protein A8 (calgranulin A) ; calgranulin A; calgranulin-A; calprotectin L1 L subunit; cystic fibrosis antigen; leukocyte L1 complex light chain; migration inhibitory factor-related protein 8; protein S100-A8; urinary stone protein band A) is found on chromosome 1 (1 q21).
  • S100A8 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 5, and which would comprise various variants from: a) acid molecules nucleic encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 5,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 5, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the S100A8 protein.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 6.
  • the LRRN3 or leucine rich repeat neuronal 3 gene (Nbla10363, FIGLER5, NLRR-3, NLRR3, fibronectin type III, immunoglobulin and leucine rich repeat domains 5; leucine-rich repeat neuronal protein 3; leucine-rich repeat protein, neuronal 3; Neuronal leucine-rich repeat protein 3) is found on chromosome 7 (7q31 .1).
  • LRRN3 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 7, and which would comprise various variants from: a) nucleic acid molecules encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 7,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 7, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the LRRN3 protein.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 8.
  • CLEC4D or C-type lectin domain family 4, member D, (MCL; MPCL; CLEC6; CLEC-6; CLECSF8) gene is found on chromosome 12 (12p13.31).
  • CLEC4D is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 9, and which would comprise various variants from: a) acid molecules nucleic encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 9,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 9, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the CLEC4D protein.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 10.
  • VNN1 or vanin 1 gene (HDLCQ8, Tiff66; CLECSF8, pantetheinase; pantetheine hydrolase; vanin-1; vannin 1; vascular non-inflammatory molecule 1) is found on chromosome 6 (6q23-q24).
  • VNN1 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 1 1, and which would comprise various variants from: a) molecules of nucleic acid encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 1,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 1 1, and wherein the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the VNN1 protein.
  • nucleic acid molecules is the collection in SEQ ID NO: 12.
  • the F5 gene or coagulation factor V (proaccelerin, labile factor), (FVL, PCCF, RPRGL1, THPH2, activated protein c cofactor; coagulation factor V; coagulation factor V jinjiang A2 domain; factor V Leiden; proaccelerin, labile factor) is found on chromosome 1 (1 q23).
  • F5 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 13, and which would comprise various variants from: a) acid molecules nucleic encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 13,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 13, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the F5 protein.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 14.
  • the IRAK3 or interleukin-1 receptor-associated kinase 3 gene, (ASRT5, IRAKM. IL-1 receptor-associated kinase M) is found on chromosome 12 (12q14.3).
  • IRAK3 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 15, and which would comprise various variants from: a) acid molecules nucleic encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 15,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 15, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the IRAK3 protein.
  • nucleic acid molecules is the collection in SEQ ID NO: 16.
  • the FAIM3 or Fas apoptotic inhibitory molecule 3 gene (FCMR, TOSO, Fe mu receptor; IgM Fe receptor; fas apoptotic inhibitory molecule 3; immunoglobulin mu Fe receptor; regulator of Fas-induced apoptosis Toso) is found on chromosome 1 (1 q32.1).
  • FAIM3 is also defined by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the protein collected in SEQ ID NO: 17, and which would comprise various variants from: a) acid molecules nucleic encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 17,
  • nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a) are nucleic acid molecules whose complementary hybrid chain with the polynucleotide sequence of a),
  • nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code
  • nucleic acid molecules encoding a polypeptide comprising the amino acid sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with SEQ ID NO: 17, and in which the polypeptide encoded by said nucleic acids possesses the activity and structural characteristics of the protein FAIM3.
  • nucleic acid molecules is the collection in the sequence SEQ ID NO: 18.
  • diagnosis refers to the ability to discriminate between individuals affected or not by pancreatic adenacarcinoma.
  • prognosis is understood as the expected evolution of a disease and refers to the assessment of the probability according to which a subject suffers from a disease as well as the assessment of its onset, developmental status, evolution, or its regression, and / or the prognosis of the course of the disease in the future. As those skilled in the art will understand, such assessment, although preferred, may not be correct for 100% of the subjects to be diagnosed. The term, however, requires that a statistically significant part of the subjects can be identified as having the disease or having a predisposition to it.
  • a part is statistically significant, it can be determined by the person skilled in the art using several well-known statistical evaluation tools, for example, determination of confidence intervals, determination of p-values, Student's t-test, Mann-Whitney test , etc.
  • Preferred confidence intervals are at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%.
  • P values are preferably 0.2, 0.1, 0.05.
  • Prediction of the response means, in the context of the present invention, the determination of the probability that the patient responds favorably or unfavorably to a particular therapy or treatment, including surgical treatment.
  • prediction refers to an individual evaluation of any parameter that may be useful in determining the evolution of a patient.
  • the prediction of the clinical response to treatment although it is preferred, does not need to be correct for 100% of the subjects to be diagnosed or evaluated. The term, however, requires that a statistically significant part of the subjects can be identified as having an increased probability of having a positive response.
  • Intervals Preferred confidence are at least 50%>, at least 60%>, at least 70%>, at least 80%>, at least 90%), at least 95%>.
  • P values are preferably 0.2, 0.1 or 0.05.
  • the prediction of the clinical response can be made using any assessment criteria used in oncology and known to the person skilled in the art.
  • the confidence intervals are at least 90%, at least 95%, at least 97%, at least 98% or at least 99%.
  • the value of p is less than 0.1, 0.05, 0.01, 0.005 or 0.0001.
  • the present invention makes it possible to correctly detect the disease differentially by at least 60%, more preferably at least 70%, much more preferably at least 80%, or even much more preferably at least 90%. of the subjects of a certain group or population analyzed.
  • an "isolated biological sample” includes, but is not limited to, cells, tissues and / or biological fluids of an organism, obtained by any method known to a person skilled in the art.
  • the isolated biological sample comprises peripheral blood mononuclear cells PBMCs.
  • peripheral blood mononuclear cell is understood as a blood cell characterized by having a single round nucleus, such as lymphocytes, monocytes or macrophages. These blood cells are a critical component in the immune system, specifically to combat infections
  • the lymphocyte population consists of T cells (CD4 and CD8 positive ⁇ 75%).
  • These cells are often obtained from the blood using fi lcol, a hydrophilic polysaccharide that separates layers of blood, with monocytes and lymphocytes forming a buffy coat under the plasma layer.
  • This buffy contains the PBMCs.
  • extraction methods known in the state of the art, such as, for example, extracting them from whole blood with a hypotonic lysis solution that preferably smooths red blood cells. This method gives rise to neutrophils and other polymorphonuclear cells (PMN) important in innate immune defense.
  • PMN polymorphonuclear cells
  • the term “individual” is not intended to be limiting in any aspect, and may be of any age, sex and physical condition.
  • the reference amount is obtained from the constitutive expression values of the genes, in a group of healthy individuals, or from the expression of the genes in the group of individuals before being subjected to treatment.
  • the reference amount will be, for example, in the case of differentiation between patients affected by pancreatic adenocarcinoma of healthy individuals, the constitutive expression of the gene in a control group of healthy individuals.
  • the control group will consist of a group of patients with adenocarcinoma of the pancreas that did not have that clinical manifestation.
  • reference samples can be obtained from individuals at different stages of treatment.
  • the sample or reference samples can be, for example, obtained from the blood cells of a patient with adenocarcinoma of the pancreas, in a certain clinical phase.
  • the reference amount is obtained from a reference sample.
  • the reference amount can also be obtained, for example, from the limits of normal distribution of an amount found in samples obtained from a population of individuals with adenocarcinoma of the pancreas in different phases, by statistical techniques well known.
  • the reference sample is obtained from patients before and after treatment.
  • Expression levels of one or more of these genes may be indicative of a subject's response or lack of response to treatment.
  • the answer is a reduction in tumor burden.
  • the response is a clinical result, which implies an improvement in the patient's condition or absence of deterioration.
  • the response is progression-free survival or an increase in overall survival.
  • CR Full Response
  • Partial Response At least a 30% decrease in the sum of diameters of the target lesions, taking as reference the initial value of the sum of diameters.
  • Progressive Disease At least a 20% increase in the sum of diameters of the target lesions, taking as a reference the smallest sum in the study (this includes the initial value of the sum if this is the smallest of the study). In addition to the relative increase of 20%, the sum must also demonstrate an absolute increase of at least 5 mm. (Note: the appearance of one or more new lesions is also considered progression).
  • Stable Disease Neither a sufficient decrease to qualify as PR nor a sufficient increase to qualify as PD, taking as reference the smallest sum of diameters during the study.
  • the terms CR, PR, PD and SD are defined in accordance with the above definitions 1 to 4 taken from the revised RECIST Guidelines of Eisenhauer et al., Eur. J. Cancer, 45 (2009) 228-247.
  • the treatment comprises the administration of Gemcitabine associated with Erlotinib. More preferably, the treatment comprises the administration of intravenous Gemcitabine 1, 000 mg / m2, days 1, 8, and 15 4-week cycles associated with continuous daily oral Erlotinib 100 mg. The treatment will be administered until documented progression of the disease or unacceptable toxicity.
  • Another aspect of the invention relates to a method for monitoring the evolution of pancreatic adenocarcinoma in an individual, hereinafter third method of the invention, comprising performing at least twice the sequence of steps (a) - ( b) according to the first or second method of the invention, in biological samples from the same individual, and isolated at different times.
  • Another aspect of the invention relates to a pharmaceutical composition
  • a pharmaceutical composition comprising a modulating agent of at least one of the genes that are selected from ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3, and / or FAIM3, for Treat an individual diagnosed with pancreatic adenocarcinoma according to the methods of the invention.
  • the modulating agent is an inhibitor of the expression of said genes.
  • Another aspect of the invention relates to an antibody for treating an individual diagnosed with pancreatic adenocarcinoma, identifiable by a method of the invention, wherein the antibody is selected from among anti-ANKRD22, anti-ARG1, anti-S100A8, anti-LRRN3 , anti-CLEC4D, anti-VNN1, anti-F5, anti-IRAK3, and / or anti-FAIM3.
  • kit or device of the invention comprising the elements necessary to analyze the level of expression of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1 genes , F5, IRAK3, and / or FAIM3.
  • the kit may contain oligonucleotides designed from a known sequence or an mRNA of the genes, and / or capable of hybridizing with the sequence of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, and / or FAIM3 genes, for subsequent amplification by PCR.
  • the sequence of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3 and / or FAIM3 genes is the nucleotide sequence SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16 and / or SEQ ID NO: 18, respectively.
  • the kit or device of the invention comprises TaqMan probes specific to Applied Biosystems ANKRD22: Ref. Hs00944018_m1; ARG1: Ref. Hs00968979_m1; S100A8: Ref. Hs00374264_g1; LRRN3: Ref. Hs00539582_s1; CLEC4D: Ref. Hs00431023_m1; VNN1: Ref. Hs01546812_m1; F5: Ref. Hs00914120_m1
  • IRAK3 Ref. Hs00936103_m1
  • FAIM3 Ref. Hs00193770_m1.
  • step (b) More preferably it comprises the means necessary to compare the amount detected in step (b) with a reference amount.
  • Said kit may contain all those reagents necessary to analyze the level of expression of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, and / or FAIM3 genes by any of the methods described hereinbefore.
  • the kit can also include, without any limitation, buffers, agents to prevent contamination, inhibitors of protein degradation, etc.
  • the kit can include all the supports and containers necessary for commissioning and optimization.
  • the kit further comprises instructions for carrying out any of the methods of the invention.
  • kits suitable for RQ-PCR a technique for quantifying sensitive and reproducible gene expression
  • the kit additionally comprises a polyT oligonucleotide primer in addition to the kit oligonucleotide (s).
  • oligonucleotide oligonucleotide (s) (hybridization probe) complementary to (part of) the target sequence of the RNA of interest.
  • the kit comprises a microarray, or microarray of the invention.
  • An RNA microarray is a matrix on a solid substrate (usually a glass holder or a cell of a thin silicon film) that evaluates large amounts of different RNAs that are detectable by specific probes immobilized on spots on a solid substrate.
  • Each spot contains a specific nucleic acid sequence, usually a DNA sequence, such as probes (or indicators).
  • the number of spots is not limited in any way, there is a preferred embodiment in which the microarray is customized for the methods of the invention.
  • said custom matrix comprises fifty spots or less, such as thirty spots or less, including twenty spots or less. Therefore, another aspect of the invention relates to a microarray comprising oligonucleotides designed from a known sequence or an mRNA of the genes, and / or capable of hybridizing with the sequence of the ANKRD22, ARG1, S100A8, LRRN3 genes, CLEC4D, VNN1, F5, IRAK3, and / or FAIM3 More preferably, the sequence of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, ⁇ RAK3 and / or FAIM3 genes is the nucleotide sequence SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16 and / or SEQ ID NO: 18, respectively.
  • microarray of the invention comprising oligonucleotides or single channel microarrays designed from a known sequence or an mRNA of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D genes. , VNN1, F5, ⁇ RAK3, and / or FAIM3.
  • the sequence of the ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3 and / or FAIM3 genes is the nucleotide sequence SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16 and / or SEQ ID NO: 18, respectively.
  • oligonucleotide sequences are constructed on the surface of a chip by sequential elongation of a growing chain with a single nucleotide using photolithography.
  • oligonucleotides are anchored by the 3 'end by a method of selective activation of nucleotides, protected by a photolabile reagent, by the selective incidence of light through a photomask.
  • the photomask can be physical or virtual.
  • the oligonucleotide probes can be between 10 and 100 nucleotides, more preferably, between 20 and 70 nucleotides, and even more preferably, between 24 and 30 nucleotides.
  • oligonucleotides per gene preferably about 40 oligonucleotides per gene are used.
  • Synthesis in situ on a solid support could be done using ink-jet technology, which requires longer probes.
  • the supports could be, but not limited to, filters or membranes of NC or nylon (charged), silicon, or glass slides for microscopes covered with aminosilanes, polylysine, aldehydes or epoxy.
  • the probe is each of the chip samples.
  • the target is the sample to be analyzed: messenger RNA, total RNA, a PCR fragment, etc.
  • kits or devices for obtaining data useful in the diagnosis, prognosis or response to the treatment of pancreatic adenocarcinoma.
  • Another aspect of the invention relates to a computer-readable storage medium comprising program instructions capable of having a computer perform the steps of any of the methods of the invention (of the first or second method of the invention ).
  • Another aspect of the invention relates to a transmissible signal comprising program instructions capable of having a computer perform the steps of any of the methods of the invention.
  • polynucleotide and “nucleic acid” are used interchangeably herein, referring to polymeric forms of nucleotides of any length, both ribonucleotides (RNA or RNA) and deoxyribonucleotides (DNA or DNA).
  • amino acid sequence a polymeric form of amino acids of any length, which may be coding or non-coding, Chemically or biochemically modified.
  • peripheral blood samples 12 ml of peripheral blood were collected from patients and control subjects in PAXgene tubes (PreAnakytix GmbH, Switzerland). Blood samples were collected at the Virgen de las Nieves Hospital (Granada) between January 2009, July 2012. Peripheral blood samples were left 24 hours at room temperature for stabilization.
  • pancreatic cancer ⁇ Pancreatic Ductal Adenocarcinoma PDAC
  • pancreatic cancer ⁇ Pancreatic Ductal Adenocarcinoma PDAC
  • RNA from PBMCs Isolation of total RNA from PBMCs
  • PAXgene ® Blood RNA system PreAnalytix GmbH, Switzerland
  • the protocol followed to extract the RNA from the collected blood cells was the protocol recommended by the manufacturer, without modifications.
  • RNA concentration and integrity was measured using a NanoDrop 2000c spectrophotometer (Thermo Scientific Wilmington, USA).
  • Hybridizations were carried out in whole human genome microarrays to identify potential PDAC markers.
  • Microarray hybridization, scanning and analysis were carried out in the Genomics Unit of the Andalusian Center for Molecular Biology and Regenerative Medicine (CABIMER, Sevilla).
  • Gene expression profiles of PBMC samples of patients with PDAC and healthy controls were carried out in Affymetrix microarrays. Briefly, 1 g of high quality total RNA was used to synthesize double stranded cDNA and then biotin-labeled cRNA was produced. The resulting biotin-labeled cRNA is recovered and purified and then the chips are hybridized. After being washed and dyed, the matrices (arrays) were scanned using a Gene-Chip Scanner 3000 7G (Affymetrix Inc).
  • the Log2 transformation was applied to all proportions, with data normalization. After normalization, the 40 subjects (20 with PDAC and 20 healthy controls) closest to normal values were selected.
  • RNA sample obtained from each RNA sample obtained.
  • the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems) was used for reverse transcriptase PCR. Briefly, 1 g of total RNA was reverse transcribed using the MultiScribe TM reverse transcriptase according to the manufacturer's instructions, in a final volume of 20 ml. The program was as follows: 25 ° C 10 min, 37 ° C 120 min, 85 ° C 5nin.
  • the Q-RT-PCR was carried out from each cDNA sample. Each sample was run in quadruplicate and the GAPDH gene was used to normalize the results.
  • the cDNA was mixed with TaqMan Fast PCR Universal Master Mix (Applied Biosystems) and with Taqman ® (Applied Biosystems) probes and primers corresponding to the previously selected genes in a reaction volume of 20 ⁇ .
  • VNN1 was the second that presented the greatest increase
  • RQ is the number of times the gene expression is increased compared to the control sample (healthy controls). Each sample is compared with the same control.

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Abstract

L'invention concerne une méthode d'obtention de données utiles pour le diagnostic précoce de l'adénocarcinome du pancréas, ainsi que pour évaluer la réponse au traitement de ladite maladie, kit et utilisations de ce dernier basé sur l'expression des gènes ANKRD22, ARG1, S100A8, LRRN3, CLEC4D, VNN1, F5, IRAK3, et/ou FAIM3.
PCT/ES2013/070786 2012-11-14 2013-11-12 Méthode d'obtention de données utiles pour le diagnostic, le pronostic et la prédiction d'une réponse au traitement de l'adénocarcinome du pancréas WO2014076342A1 (fr)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020254667A1 (fr) 2019-06-19 2020-12-24 Universidad De Granada Séries de métabolites utilisées en tant que biomarqueurs pour le diagnostic du cancer du pancréas
WO2021052049A1 (fr) * 2019-09-17 2021-03-25 浙江大学 Utilisation de ankrd22 comme cible dans la préparation d'un agent protecteur pour la réparation de la muqueuse gastro-intestinale

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BAINE MICHAEL J ET AL.: "Transcriptional profiling of peripheral blood mononuclear cells in pancreatic cancer patients identifies novel genes with potential diagnostic utility..", PLOS ONE UNITED STATES, vol. 6, no. 2, 30 November 2010 (2010-11-30), pages e17014, ISSN: 1932-6203 *
HUANG HAI ET AL.: "Novel blood biomarkers of pancreatic cancer-associated diabetes mellitus identified by peripheral blood-based gene expression profiles..", THE AMERICAN JOURNAL OF GASTROENTEROLOGY UNITED STATES, vol. 105, no. 7, 30 June 2010 (2010-06-30), pages 1661 - 1669, ISSN: 1572-0241 *
SABBAGHIAN M SHIRIN ET AL.: "Levels of Elevated Circulating Endothelial Cell Decline after Tumor Resection in Patients with Pancreatic Ductal Adenocarcinoma.", ANTICANCER RESEARCH, vol. 30, no. 7, 30 June 2010 (2010-06-30), pages 2911 - 2917, ISSN: 0250-7005 *
ZHOU JIAHUA ET AL.: "Marker expression in circulating cancer cells of pancreatic cancer patients..", THE JOURNAL OF SURGICAL RESEARCH UNITED STATES, vol. 171, no. 2, 30 November 2011 (2011-11-30), pages 631 - 636, ISSN: 1095-8673 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020254667A1 (fr) 2019-06-19 2020-12-24 Universidad De Granada Séries de métabolites utilisées en tant que biomarqueurs pour le diagnostic du cancer du pancréas
WO2021052049A1 (fr) * 2019-09-17 2021-03-25 浙江大学 Utilisation de ankrd22 comme cible dans la préparation d'un agent protecteur pour la réparation de la muqueuse gastro-intestinale

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